CA2971213C - Procedes et compositions pour modification genetique ciblee par ciblage multiple en une seule etape - Google Patents
Procedes et compositions pour modification genetique ciblee par ciblage multiple en une seule etape Download PDFInfo
- Publication number
- CA2971213C CA2971213C CA2971213A CA2971213A CA2971213C CA 2971213 C CA2971213 C CA 2971213C CA 2971213 A CA2971213 A CA 2971213A CA 2971213 A CA2971213 A CA 2971213A CA 2971213 C CA2971213 C CA 2971213C
- Authority
- CA
- Canada
- Prior art keywords
- nucleic acid
- ltvec
- acid insert
- cell
- sequence
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
- 238000000034 method Methods 0.000 title claims abstract description 413
- 230000008685 targeting Effects 0.000 title claims abstract description 362
- 239000000203 mixture Substances 0.000 title abstract description 38
- 238000012239 gene modification Methods 0.000 title abstract description 31
- 230000005017 genetic modification Effects 0.000 title abstract description 28
- 235000013617 genetically modified food Nutrition 0.000 title abstract description 28
- 150000007523 nucleic acids Chemical class 0.000 claims abstract description 799
- 102000039446 nucleic acids Human genes 0.000 claims abstract description 745
- 108020004707 nucleic acids Proteins 0.000 claims abstract description 745
- 239000013598 vector Substances 0.000 claims abstract description 253
- 241000282414 Homo sapiens Species 0.000 claims abstract description 210
- 101710163270 Nuclease Proteins 0.000 claims abstract description 188
- 239000003795 chemical substances by application Substances 0.000 claims abstract description 120
- 238000002744 homologous recombination Methods 0.000 claims abstract description 82
- 230000006801 homologous recombination Effects 0.000 claims abstract description 82
- 210000004027 cell Anatomy 0.000 claims description 594
- 108090000623 proteins and genes Proteins 0.000 claims description 214
- 108020004414 DNA Proteins 0.000 claims description 118
- 108020005004 Guide RNA Proteins 0.000 claims description 111
- 102000004169 proteins and genes Human genes 0.000 claims description 103
- 102000040430 polynucleotide Human genes 0.000 claims description 95
- 108091033319 polynucleotide Proteins 0.000 claims description 95
- 239000002157 polynucleotide Substances 0.000 claims description 95
- 230000010354 integration Effects 0.000 claims description 74
- 238000012217 deletion Methods 0.000 claims description 71
- 230000037430 deletion Effects 0.000 claims description 71
- 108010017070 Zinc Finger Nucleases Proteins 0.000 claims description 67
- 108091028043 Nucleic acid sequence Proteins 0.000 claims description 65
- 210000001161 mammalian embryo Anatomy 0.000 claims description 63
- 210000004962 mammalian cell Anatomy 0.000 claims description 59
- 230000001105 regulatory effect Effects 0.000 claims description 59
- 230000006798 recombination Effects 0.000 claims description 55
- 238000005215 recombination Methods 0.000 claims description 55
- 230000005782 double-strand break Effects 0.000 claims description 53
- 108091033409 CRISPR Proteins 0.000 claims description 52
- 241000894007 species Species 0.000 claims description 39
- 241000283984 Rodentia Species 0.000 claims description 38
- 108700028369 Alleles Proteins 0.000 claims description 35
- 239000003550 marker Substances 0.000 claims description 28
- 210000005260 human cell Anatomy 0.000 claims description 27
- 230000035772 mutation Effects 0.000 claims description 25
- 230000005783 single-strand break Effects 0.000 claims description 24
- 238000003780 insertion Methods 0.000 claims description 23
- 230000037431 insertion Effects 0.000 claims description 23
- 238000010459 TALEN Methods 0.000 claims description 21
- 210000002950 fibroblast Anatomy 0.000 claims description 16
- 210000000130 stem cell Anatomy 0.000 claims description 16
- 210000003958 hematopoietic stem cell Anatomy 0.000 claims description 13
- 210000000349 chromosome Anatomy 0.000 claims description 12
- 230000001537 neural effect Effects 0.000 claims description 12
- 238000000338 in vitro Methods 0.000 claims description 10
- 108700008625 Reporter Genes Proteins 0.000 claims description 8
- 210000004263 induced pluripotent stem cell Anatomy 0.000 claims description 4
- 230000002708 enhancing effect Effects 0.000 abstract description 8
- 102000053602 DNA Human genes 0.000 description 110
- 241000699666 Mus <mouse, genus> Species 0.000 description 98
- 235000018102 proteins Nutrition 0.000 description 98
- 239000002773 nucleotide Substances 0.000 description 92
- 125000003729 nucleotide group Chemical group 0.000 description 88
- 230000004048 modification Effects 0.000 description 69
- 238000012986 modification Methods 0.000 description 69
- 241000700159 Rattus Species 0.000 description 54
- 238000003556 assay Methods 0.000 description 48
- 238000003776 cleavage reaction Methods 0.000 description 38
- 230000007017 scission Effects 0.000 description 37
- YQYJSBFKSSDGFO-UHFFFAOYSA-N Epihygromycin Natural products OC1C(O)C(C(=O)C)OC1OC(C(=C1)O)=CC=C1C=C(C)C(=O)NC1C(O)C(O)C2OCOC2C1O YQYJSBFKSSDGFO-UHFFFAOYSA-N 0.000 description 36
- 230000000295 complement effect Effects 0.000 description 35
- 239000012634 fragment Substances 0.000 description 34
- 239000000523 sample Substances 0.000 description 34
- 241001465754 Metazoa Species 0.000 description 29
- 238000012216 screening Methods 0.000 description 23
- 230000027455 binding Effects 0.000 description 22
- 229920002477 rna polymer Polymers 0.000 description 21
- 108091028113 Trans-activating crRNA Proteins 0.000 description 20
- 238000007792 addition Methods 0.000 description 19
- 241000699800 Cricetinae Species 0.000 description 17
- 241000124008 Mammalia Species 0.000 description 17
- 238000009396 hybridization Methods 0.000 description 15
- 108020004705 Codon Proteins 0.000 description 14
- -1 Sox I Proteins 0.000 description 14
- 210000001519 tissue Anatomy 0.000 description 14
- 238000010453 CRISPR/Cas method Methods 0.000 description 13
- 108091026890 Coding region Proteins 0.000 description 13
- 230000001965 increasing effect Effects 0.000 description 13
- 108090000765 processed proteins & peptides Proteins 0.000 description 13
- 150000001413 amino acids Chemical class 0.000 description 12
- 230000000694 effects Effects 0.000 description 12
- 230000001939 inductive effect Effects 0.000 description 12
- 229920001184 polypeptide Polymers 0.000 description 12
- 102000004196 processed proteins & peptides Human genes 0.000 description 12
- 230000004568 DNA-binding Effects 0.000 description 11
- 210000000472 morula Anatomy 0.000 description 11
- 210000000287 oocyte Anatomy 0.000 description 11
- 108010008532 Deoxyribonuclease I Proteins 0.000 description 10
- 102000007260 Deoxyribonuclease I Human genes 0.000 description 10
- 210000002459 blastocyst Anatomy 0.000 description 10
- 238000006467 substitution reaction Methods 0.000 description 10
- 235000001014 amino acid Nutrition 0.000 description 9
- 210000003527 eukaryotic cell Anatomy 0.000 description 9
- 230000012447 hatching Effects 0.000 description 9
- 238000010348 incorporation Methods 0.000 description 9
- 238000003753 real-time PCR Methods 0.000 description 9
- 102000004533 Endonucleases Human genes 0.000 description 8
- 108010042407 Endonucleases Proteins 0.000 description 8
- 241000193996 Streptococcus pyogenes Species 0.000 description 8
- 108091023040 Transcription factor Proteins 0.000 description 8
- 102000040945 Transcription factor Human genes 0.000 description 8
- 238000006243 chemical reaction Methods 0.000 description 8
- 230000001404 mediated effect Effects 0.000 description 8
- 239000013612 plasmid Substances 0.000 description 8
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 7
- 102000040650 (ribonucleotides)n+m Human genes 0.000 description 7
- 238000013459 approach Methods 0.000 description 7
- 210000004436 artificial bacterial chromosome Anatomy 0.000 description 7
- 230000002441 reversible effect Effects 0.000 description 7
- 238000013518 transcription Methods 0.000 description 7
- 230000035897 transcription Effects 0.000 description 7
- 241000282693 Cercopithecidae Species 0.000 description 6
- 108091092566 Extrachromosomal DNA Proteins 0.000 description 6
- 238000009015 Human TaqMan MicroRNA Assay kit Methods 0.000 description 6
- 108010077850 Nuclear Localization Signals Proteins 0.000 description 6
- 108091005461 Nucleic proteins Proteins 0.000 description 6
- 108700026244 Open Reading Frames Proteins 0.000 description 6
- 229940024606 amino acid Drugs 0.000 description 6
- 210000001106 artificial yeast chromosome Anatomy 0.000 description 6
- 210000001671 embryonic stem cell Anatomy 0.000 description 6
- 108020004999 messenger RNA Proteins 0.000 description 6
- 210000004940 nucleus Anatomy 0.000 description 6
- 230000002103 transcriptional effect Effects 0.000 description 6
- 238000001890 transfection Methods 0.000 description 6
- 230000009261 transgenic effect Effects 0.000 description 6
- 108060003951 Immunoglobulin Proteins 0.000 description 5
- 102000012960 Immunoglobulin kappa-Chains Human genes 0.000 description 5
- 108010090227 Immunoglobulin kappa-Chains Proteins 0.000 description 5
- 101100096028 Mus musculus Smok1 gene Proteins 0.000 description 5
- 241000699670 Mus sp. Species 0.000 description 5
- 108010021466 Mutant Proteins Proteins 0.000 description 5
- 102000008300 Mutant Proteins Human genes 0.000 description 5
- 108010073062 Transcription Activator-Like Effectors Proteins 0.000 description 5
- 238000004458 analytical method Methods 0.000 description 5
- 230000015572 biosynthetic process Effects 0.000 description 5
- 238000004520 electroporation Methods 0.000 description 5
- 239000003623 enhancer Substances 0.000 description 5
- 230000004927 fusion Effects 0.000 description 5
- 210000004602 germ cell Anatomy 0.000 description 5
- 102000018358 immunoglobulin Human genes 0.000 description 5
- 230000007115 recruitment Effects 0.000 description 5
- 108010054624 red fluorescent protein Proteins 0.000 description 5
- 239000000126 substance Substances 0.000 description 5
- 108091006107 transcriptional repressors Proteins 0.000 description 5
- 238000012546 transfer Methods 0.000 description 5
- 108091005957 yellow fluorescent proteins Proteins 0.000 description 5
- LRFVTYWOQMYALW-UHFFFAOYSA-N 9H-xanthine Chemical compound O=C1NC(=O)NC2=C1NC=N2 LRFVTYWOQMYALW-UHFFFAOYSA-N 0.000 description 4
- 241000283690 Bos taurus Species 0.000 description 4
- 238000011740 C57BL/6 mouse Methods 0.000 description 4
- 238000010356 CRISPR-Cas9 genome editing Methods 0.000 description 4
- 102000004190 Enzymes Human genes 0.000 description 4
- 108090000790 Enzymes Proteins 0.000 description 4
- 108010043121 Green Fluorescent Proteins Proteins 0.000 description 4
- 102000004144 Green Fluorescent Proteins Human genes 0.000 description 4
- 108010091358 Hypoxanthine Phosphoribosyltransferase Proteins 0.000 description 4
- 102000018251 Hypoxanthine Phosphoribosyltransferase Human genes 0.000 description 4
- 102100025169 Max-binding protein MNT Human genes 0.000 description 4
- 229930193140 Neomycin Natural products 0.000 description 4
- 241001494479 Pecora Species 0.000 description 4
- HCHKCACWOHOZIP-UHFFFAOYSA-N Zinc Chemical compound [Zn] HCHKCACWOHOZIP-UHFFFAOYSA-N 0.000 description 4
- 125000000539 amino acid group Chemical group 0.000 description 4
- 238000010367 cloning Methods 0.000 description 4
- 230000006378 damage Effects 0.000 description 4
- 201000010099 disease Diseases 0.000 description 4
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 4
- 238000002474 experimental method Methods 0.000 description 4
- 108010021843 fluorescent protein 583 Proteins 0.000 description 4
- 108091006047 fluorescent proteins Proteins 0.000 description 4
- 102000034287 fluorescent proteins Human genes 0.000 description 4
- 108020001507 fusion proteins Proteins 0.000 description 4
- 102000037865 fusion proteins Human genes 0.000 description 4
- 238000010362 genome editing Methods 0.000 description 4
- 238000003205 genotyping method Methods 0.000 description 4
- 239000000178 monomer Substances 0.000 description 4
- 229960004927 neomycin Drugs 0.000 description 4
- 230000008569 process Effects 0.000 description 4
- 210000001082 somatic cell Anatomy 0.000 description 4
- 239000000758 substrate Substances 0.000 description 4
- 229910052725 zinc Inorganic materials 0.000 description 4
- 239000011701 zinc Substances 0.000 description 4
- 241000271566 Aves Species 0.000 description 3
- 241000282472 Canis lupus familiaris Species 0.000 description 3
- 241000283707 Capra Species 0.000 description 3
- 241000282994 Cervidae Species 0.000 description 3
- 108091005941 EBFP Proteins 0.000 description 3
- 241000282326 Felis catus Species 0.000 description 3
- 241000287828 Gallus gallus Species 0.000 description 3
- 108700028146 Genetic Enhancer Elements Proteins 0.000 description 3
- 101000634835 Homo sapiens M1-specific T cell receptor alpha chain Proteins 0.000 description 3
- 101000634836 Homo sapiens T cell receptor alpha chain MC.7.G5 Proteins 0.000 description 3
- 102100029450 M1-specific T cell receptor alpha chain Human genes 0.000 description 3
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 3
- 241000700584 Simplexvirus Species 0.000 description 3
- 238000002105 Southern blotting Methods 0.000 description 3
- 239000004098 Tetracycline Substances 0.000 description 3
- 241000251539 Vertebrata <Metazoa> Species 0.000 description 3
- 235000004279 alanine Nutrition 0.000 description 3
- 230000004075 alteration Effects 0.000 description 3
- 230000000692 anti-sense effect Effects 0.000 description 3
- 230000001580 bacterial effect Effects 0.000 description 3
- 108091005948 blue fluorescent proteins Proteins 0.000 description 3
- 230000008859 change Effects 0.000 description 3
- 235000013330 chicken meat Nutrition 0.000 description 3
- 238000007796 conventional method Methods 0.000 description 3
- 108010082025 cyan fluorescent protein Proteins 0.000 description 3
- 210000003981 ectoderm Anatomy 0.000 description 3
- 210000002257 embryonic structure Anatomy 0.000 description 3
- 210000001900 endoderm Anatomy 0.000 description 3
- 108010048367 enhanced green fluorescent protein Proteins 0.000 description 3
- 210000001654 germ layer Anatomy 0.000 description 3
- 239000005090 green fluorescent protein Substances 0.000 description 3
- UYTPUPDQBNUYGX-UHFFFAOYSA-N guanine Chemical compound O=C1NC(N)=NC2=C1N=CN2 UYTPUPDQBNUYGX-UHFFFAOYSA-N 0.000 description 3
- HNDVDQJCIGZPNO-UHFFFAOYSA-N histidine Natural products OC(=O)C(N)CC1=CN=CN1 HNDVDQJCIGZPNO-UHFFFAOYSA-N 0.000 description 3
- 210000000688 human artificial chromosome Anatomy 0.000 description 3
- 238000002844 melting Methods 0.000 description 3
- 230000008018 melting Effects 0.000 description 3
- 210000003716 mesoderm Anatomy 0.000 description 3
- 108091070501 miRNA Proteins 0.000 description 3
- 239000002679 microRNA Substances 0.000 description 3
- 210000003470 mitochondria Anatomy 0.000 description 3
- 210000001778 pluripotent stem cell Anatomy 0.000 description 3
- 210000001236 prokaryotic cell Anatomy 0.000 description 3
- 238000012207 quantitative assay Methods 0.000 description 3
- 108091008146 restriction endonucleases Proteins 0.000 description 3
- 150000003839 salts Chemical class 0.000 description 3
- 229910052594 sapphire Inorganic materials 0.000 description 3
- 239000010980 sapphire Substances 0.000 description 3
- 239000000243 solution Substances 0.000 description 3
- 229960002180 tetracycline Drugs 0.000 description 3
- 229930101283 tetracycline Natural products 0.000 description 3
- 235000019364 tetracycline Nutrition 0.000 description 3
- 150000003522 tetracyclines Chemical class 0.000 description 3
- 210000005253 yeast cell Anatomy 0.000 description 3
- 210000004340 zona pellucida Anatomy 0.000 description 3
- YMHOBZXQZVXHBM-UHFFFAOYSA-N 2,5-dimethoxy-4-bromophenethylamine Chemical compound COC1=CC(CCN)=C(OC)C=C1Br YMHOBZXQZVXHBM-UHFFFAOYSA-N 0.000 description 2
- FWMNVWWHGCHHJJ-SKKKGAJSSA-N 4-amino-1-[(2r)-6-amino-2-[[(2r)-2-[[(2r)-2-[[(2r)-2-amino-3-phenylpropanoyl]amino]-3-phenylpropanoyl]amino]-4-methylpentanoyl]amino]hexanoyl]piperidine-4-carboxylic acid Chemical compound C([C@H](C(=O)N[C@H](CC(C)C)C(=O)N[C@H](CCCCN)C(=O)N1CCC(N)(CC1)C(O)=O)NC(=O)[C@H](N)CC=1C=CC=CC=1)C1=CC=CC=C1 FWMNVWWHGCHHJJ-SKKKGAJSSA-N 0.000 description 2
- KDCGOANMDULRCW-UHFFFAOYSA-N 7H-purine Chemical compound N1=CNC2=NC=NC2=C1 KDCGOANMDULRCW-UHFFFAOYSA-N 0.000 description 2
- 102000006822 Agouti Signaling Protein Human genes 0.000 description 2
- 108010072151 Agouti Signaling Protein Proteins 0.000 description 2
- 102100036475 Alanine aminotransferase 1 Human genes 0.000 description 2
- 241000272517 Anseriformes Species 0.000 description 2
- 241000894006 Bacteria Species 0.000 description 2
- 101710201279 Biotin carboxyl carrier protein Proteins 0.000 description 2
- 108010045123 Blasticidin-S deaminase Proteins 0.000 description 2
- 101150111062 C gene Proteins 0.000 description 2
- 108091079001 CRISPR RNA Proteins 0.000 description 2
- 108091005944 Cerulean Proteins 0.000 description 2
- 241000579895 Chlorostilbon Species 0.000 description 2
- 108700010070 Codon Usage Proteins 0.000 description 2
- 241000484025 Cuniculus Species 0.000 description 2
- 108091005943 CyPet Proteins 0.000 description 2
- 108010053770 Deoxyribonucleases Proteins 0.000 description 2
- 102000016911 Deoxyribonucleases Human genes 0.000 description 2
- 101001091269 Escherichia coli Hygromycin-B 4-O-kinase Proteins 0.000 description 2
- ZHNUHDYFZUAESO-UHFFFAOYSA-N Formamide Chemical compound NC=O ZHNUHDYFZUAESO-UHFFFAOYSA-N 0.000 description 2
- 102000005720 Glutathione transferase Human genes 0.000 description 2
- 108010070675 Glutathione transferase Proteins 0.000 description 2
- 101710154606 Hemagglutinin Proteins 0.000 description 2
- 241000282412 Homo Species 0.000 description 2
- 101001046587 Homo sapiens Krueppel-like factor 1 Proteins 0.000 description 2
- 101001139146 Homo sapiens Krueppel-like factor 2 Proteins 0.000 description 2
- 101001139134 Homo sapiens Krueppel-like factor 4 Proteins 0.000 description 2
- 101001139130 Homo sapiens Krueppel-like factor 5 Proteins 0.000 description 2
- 101000984042 Homo sapiens Protein lin-28 homolog A Proteins 0.000 description 2
- 108010025815 Kanamycin Kinase Proteins 0.000 description 2
- 102100022248 Krueppel-like factor 1 Human genes 0.000 description 2
- 102100020675 Krueppel-like factor 2 Human genes 0.000 description 2
- 102100020677 Krueppel-like factor 4 Human genes 0.000 description 2
- 102100020680 Krueppel-like factor 5 Human genes 0.000 description 2
- QNAYBMKLOCPYGJ-REOHCLBHSA-N L-alanine Chemical compound C[C@H](N)C(O)=O QNAYBMKLOCPYGJ-REOHCLBHSA-N 0.000 description 2
- 241000282560 Macaca mulatta Species 0.000 description 2
- 241000282341 Mustela putorius furo Species 0.000 description 2
- 108091057508 Myc family Proteins 0.000 description 2
- 102100038895 Myc proto-oncogene protein Human genes 0.000 description 2
- 101710135898 Myc proto-oncogene protein Proteins 0.000 description 2
- 208000009869 Neu-Laxova syndrome Diseases 0.000 description 2
- 108091092724 Noncoding DNA Proteins 0.000 description 2
- 241000283973 Oryctolagus cuniculus Species 0.000 description 2
- 101710093908 Outer capsid protein VP4 Proteins 0.000 description 2
- 101710135467 Outer capsid protein sigma-1 Proteins 0.000 description 2
- 241000288906 Primates Species 0.000 description 2
- 101710176177 Protein A56 Proteins 0.000 description 2
- 102100025460 Protein lin-28 homolog A Human genes 0.000 description 2
- 230000004570 RNA-binding Effects 0.000 description 2
- 101150001847 Sox15 gene Proteins 0.000 description 2
- 241000194020 Streptococcus thermophilus Species 0.000 description 2
- 101001091268 Streptomyces hygroscopicus Hygromycin-B 7''-O-kinase Proteins 0.000 description 2
- 241000187191 Streptomyces viridochromogenes Species 0.000 description 2
- 241000203587 Streptosporangium roseum Species 0.000 description 2
- 241000282887 Suidae Species 0.000 description 2
- 102000002933 Thioredoxin Human genes 0.000 description 2
- 102000006601 Thymidine Kinase Human genes 0.000 description 2
- 108020004440 Thymidine kinase Proteins 0.000 description 2
- 101710150448 Transcriptional regulator Myc Proteins 0.000 description 2
- 108700019146 Transgenes Proteins 0.000 description 2
- 241000545067 Venus Species 0.000 description 2
- 241000700605 Viruses Species 0.000 description 2
- 108010027570 Xanthine phosphoribosyltransferase Proteins 0.000 description 2
- 230000009471 action Effects 0.000 description 2
- 210000004102 animal cell Anatomy 0.000 description 2
- 230000004071 biological effect Effects 0.000 description 2
- 230000003197 catalytic effect Effects 0.000 description 2
- 230000001413 cellular effect Effects 0.000 description 2
- 210000004978 chinese hamster ovary cell Anatomy 0.000 description 2
- 102000021178 chitin binding proteins Human genes 0.000 description 2
- 108091011157 chitin binding proteins Proteins 0.000 description 2
- 210000003763 chloroplast Anatomy 0.000 description 2
- 238000005352 clarification Methods 0.000 description 2
- 238000011259 co-electroporation Methods 0.000 description 2
- 230000021615 conjugation Effects 0.000 description 2
- 230000003247 decreasing effect Effects 0.000 description 2
- 238000013461 design Methods 0.000 description 2
- 238000001514 detection method Methods 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 230000018109 developmental process Effects 0.000 description 2
- 239000010976 emerald Substances 0.000 description 2
- 229910052876 emerald Inorganic materials 0.000 description 2
- 108010050663 endodeoxyribonuclease CreI Proteins 0.000 description 2
- 108010026638 endodeoxyribonuclease FokI Proteins 0.000 description 2
- 210000002889 endothelial cell Anatomy 0.000 description 2
- 230000007159 enucleation Effects 0.000 description 2
- 239000007850 fluorescent dye Substances 0.000 description 2
- 102000054766 genetic haplotypes Human genes 0.000 description 2
- 239000000185 hemagglutinin Substances 0.000 description 2
- 229910052739 hydrogen Inorganic materials 0.000 description 2
- 239000001257 hydrogen Substances 0.000 description 2
- 238000001727 in vivo Methods 0.000 description 2
- 230000003993 interaction Effects 0.000 description 2
- 210000003292 kidney cell Anatomy 0.000 description 2
- 238000011068 loading method Methods 0.000 description 2
- 241001515942 marmosets Species 0.000 description 2
- 239000011159 matrix material Substances 0.000 description 2
- 230000003278 mimic effect Effects 0.000 description 2
- 238000010369 molecular cloning Methods 0.000 description 2
- 238000002703 mutagenesis Methods 0.000 description 2
- 231100000350 mutagenesis Toxicity 0.000 description 2
- 108091008800 n-Myc Proteins 0.000 description 2
- 210000000653 nervous system Anatomy 0.000 description 2
- 210000004498 neuroglial cell Anatomy 0.000 description 2
- 210000002569 neuron Anatomy 0.000 description 2
- 238000005457 optimization Methods 0.000 description 2
- 210000000056 organ Anatomy 0.000 description 2
- 210000001672 ovary Anatomy 0.000 description 2
- 230000036961 partial effect Effects 0.000 description 2
- 229920000642 polymer Polymers 0.000 description 2
- 238000000746 purification Methods 0.000 description 2
- RXWNCPJZOCPEPQ-NVWDDTSBSA-N puromycin Chemical compound C1=CC(OC)=CC=C1C[C@H](N)C(=O)N[C@H]1[C@@H](O)[C@H](N2C3=NC=NC(=C3N=C2)N(C)C)O[C@@H]1CO RXWNCPJZOCPEPQ-NVWDDTSBSA-N 0.000 description 2
- 108010045647 puromycin N-acetyltransferase Proteins 0.000 description 2
- 230000002829 reductive effect Effects 0.000 description 2
- 230000022532 regulation of transcription, DNA-dependent Effects 0.000 description 2
- 230000008672 reprogramming Effects 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 230000035939 shock Effects 0.000 description 2
- 150000003384 small molecules Chemical class 0.000 description 2
- 125000006850 spacer group Chemical group 0.000 description 2
- 238000012453 sprague-dawley rat model Methods 0.000 description 2
- 230000004960 subcellular localization Effects 0.000 description 2
- 238000003786 synthesis reaction Methods 0.000 description 2
- 238000010381 tandem affinity purification Methods 0.000 description 2
- 108060008226 thioredoxin Proteins 0.000 description 2
- 229940094937 thioredoxin Drugs 0.000 description 2
- 238000003151 transfection method Methods 0.000 description 2
- 238000011144 upstream manufacturing Methods 0.000 description 2
- 229940075420 xanthine Drugs 0.000 description 2
- NHBKXEKEPDILRR-UHFFFAOYSA-N 2,3-bis(butanoylsulfanyl)propyl butanoate Chemical compound CCCC(=O)OCC(SC(=O)CCC)CSC(=O)CCC NHBKXEKEPDILRR-UHFFFAOYSA-N 0.000 description 1
- 241000007910 Acaryochloris marina Species 0.000 description 1
- 241001135192 Acetohalobium arabaticum Species 0.000 description 1
- 241001464929 Acidithiobacillus caldus Species 0.000 description 1
- 241000605222 Acidithiobacillus ferrooxidans Species 0.000 description 1
- 241000251468 Actinopterygii Species 0.000 description 1
- 241001136782 Alca Species 0.000 description 1
- 102000007698 Alcohol dehydrogenase Human genes 0.000 description 1
- 108010021809 Alcohol dehydrogenase Proteins 0.000 description 1
- 102000002260 Alkaline Phosphatase Human genes 0.000 description 1
- 108020004774 Alkaline Phosphatase Proteins 0.000 description 1
- 241000190857 Allochromatium vinosum Species 0.000 description 1
- 241000147155 Ammonifex degensii Species 0.000 description 1
- 241001495180 Arthrospira Species 0.000 description 1
- 241000620196 Arthrospira maxima Species 0.000 description 1
- 240000002900 Arthrospira platensis Species 0.000 description 1
- 235000016425 Arthrospira platensis Nutrition 0.000 description 1
- 108091005950 Azurite Proteins 0.000 description 1
- 102000052609 BRCA2 Human genes 0.000 description 1
- 108700020462 BRCA2 Proteins 0.000 description 1
- 241000906059 Bacillus pseudomycoides Species 0.000 description 1
- 241000157302 Bison bison athabascae Species 0.000 description 1
- 101150008921 Brca2 gene Proteins 0.000 description 1
- 241000823281 Burkholderiales bacterium Species 0.000 description 1
- 238000010354 CRISPR gene editing Methods 0.000 description 1
- 101150018129 CSF2 gene Proteins 0.000 description 1
- 101150069031 CSN2 gene Proteins 0.000 description 1
- 101100257372 Caenorhabditis elegans sox-3 gene Proteins 0.000 description 1
- 102000000584 Calmodulin Human genes 0.000 description 1
- 108010041952 Calmodulin Proteins 0.000 description 1
- 241001496650 Candidatus Desulforudis Species 0.000 description 1
- 241000700198 Cavia Species 0.000 description 1
- 108010051109 Cell-Penetrating Peptides Proteins 0.000 description 1
- 102000020313 Cell-Penetrating Peptides Human genes 0.000 description 1
- 108091005960 Citrine Proteins 0.000 description 1
- 241000193163 Clostridioides difficile Species 0.000 description 1
- 241000193155 Clostridium botulinum Species 0.000 description 1
- 241000907165 Coleofasciculus chthonoplastes Species 0.000 description 1
- 241000699802 Cricetulus griseus Species 0.000 description 1
- 241000065716 Crocosphaera watsonii Species 0.000 description 1
- 241000159506 Cyanothece Species 0.000 description 1
- 230000004544 DNA amplification Effects 0.000 description 1
- 238000001712 DNA sequencing Methods 0.000 description 1
- 108090000626 DNA-directed RNA polymerases Proteins 0.000 description 1
- 102000004163 DNA-directed RNA polymerases Human genes 0.000 description 1
- 229920002307 Dextran Polymers 0.000 description 1
- 101100300807 Drosophila melanogaster spn-A gene Proteins 0.000 description 1
- 206010059866 Drug resistance Diseases 0.000 description 1
- 101000889905 Enterobacteria phage RB3 Intron-associated endonuclease 3 Proteins 0.000 description 1
- 101000889904 Enterobacteria phage T4 Defective intron-associated endonuclease 3 Proteins 0.000 description 1
- 101000889900 Enterobacteria phage T4 Intron-associated endonuclease 1 Proteins 0.000 description 1
- 101000889899 Enterobacteria phage T4 Intron-associated endonuclease 2 Proteins 0.000 description 1
- 241000283086 Equidae Species 0.000 description 1
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 1
- 241000206602 Eukaryota Species 0.000 description 1
- 241000326311 Exiguobacterium sibiricum Species 0.000 description 1
- 108010007577 Exodeoxyribonuclease I Proteins 0.000 description 1
- 108700024394 Exon Proteins 0.000 description 1
- 241000192016 Finegoldia magna Species 0.000 description 1
- 108700007698 Genetic Terminator Regions Proteins 0.000 description 1
- KOSRFJWDECSPRO-WDSKDSINSA-N Glu-Glu Chemical compound OC(=O)CC[C@H](N)C(=O)N[C@@H](CCC(O)=O)C(O)=O KOSRFJWDECSPRO-WDSKDSINSA-N 0.000 description 1
- 241000700721 Hepatitis B virus Species 0.000 description 1
- MAJYPBAJPNUFPV-BQBZGAKWSA-N His-Cys Chemical compound SC[C@@H](C(O)=O)NC(=O)[C@@H](N)CC1=CN=CN1 MAJYPBAJPNUFPV-BQBZGAKWSA-N 0.000 description 1
- 102000008157 Histone Demethylases Human genes 0.000 description 1
- 108010074870 Histone Demethylases Proteins 0.000 description 1
- 102000003893 Histone acetyltransferases Human genes 0.000 description 1
- 108090000246 Histone acetyltransferases Proteins 0.000 description 1
- 241001272567 Hominoidea Species 0.000 description 1
- 101000744174 Homo sapiens DNA-3-methyladenine glycosylase Proteins 0.000 description 1
- 241000713772 Human immunodeficiency virus 1 Species 0.000 description 1
- 108010067060 Immunoglobulin Variable Region Proteins 0.000 description 1
- 108091092195 Intron Proteins 0.000 description 1
- 241001430080 Ktedonobacter racemifer Species 0.000 description 1
- CKLJMWTZIZZHCS-REOHCLBHSA-N L-aspartic acid Chemical compound OC(=O)[C@@H](N)CC(O)=O CKLJMWTZIZZHCS-REOHCLBHSA-N 0.000 description 1
- 241000186673 Lactobacillus delbrueckii Species 0.000 description 1
- 241000186869 Lactobacillus salivarius Species 0.000 description 1
- 241000713666 Lentivirus Species 0.000 description 1
- 108060001084 Luciferase Proteins 0.000 description 1
- 239000005089 Luciferase Substances 0.000 description 1
- 241001134698 Lyngbya Species 0.000 description 1
- 101710175625 Maltose/maltodextrin-binding periplasmic protein Proteins 0.000 description 1
- 241000501784 Marinobacter sp. Species 0.000 description 1
- 108010063312 Metalloproteins Proteins 0.000 description 1
- 102000010750 Metalloproteins Human genes 0.000 description 1
- 241000204637 Methanohalobium evestigatum Species 0.000 description 1
- 108060004795 Methyltransferase Proteins 0.000 description 1
- 241000192710 Microcystis aeruginosa Species 0.000 description 1
- 241000190928 Microscilla marina Species 0.000 description 1
- 108010085220 Multiprotein Complexes Proteins 0.000 description 1
- 102000007474 Multiprotein Complexes Human genes 0.000 description 1
- 241000711408 Murine respirovirus Species 0.000 description 1
- 241000699660 Mus musculus Species 0.000 description 1
- 101100494762 Mus musculus Nedd9 gene Proteins 0.000 description 1
- 101100257376 Mus musculus Sox3 gene Proteins 0.000 description 1
- 241000167285 Natranaerobius thermophilus Species 0.000 description 1
- 101100385413 Neurospora crassa (strain ATCC 24698 / 74-OR23-1A / CBS 708.71 / DSM 1257 / FGSC 987) csm-3 gene Proteins 0.000 description 1
- 241000919925 Nitrosococcus halophilus Species 0.000 description 1
- 241001515112 Nitrosococcus watsonii Species 0.000 description 1
- 241000203619 Nocardiopsis dassonvillei Species 0.000 description 1
- 241001223105 Nodularia spumigena Species 0.000 description 1
- 241000192673 Nostoc sp. Species 0.000 description 1
- 241000192520 Oscillatoria sp. Species 0.000 description 1
- 238000010222 PCR analysis Methods 0.000 description 1
- 108091081548 Palindromic sequence Proteins 0.000 description 1
- 241000142651 Pelotomaculum thermopropionicum Species 0.000 description 1
- 108010088535 Pep-1 peptide Proteins 0.000 description 1
- 241000983938 Petrotoga mobilis Species 0.000 description 1
- 241000286209 Phasianidae Species 0.000 description 1
- 241000512220 Polaromonas Species 0.000 description 1
- 241001599925 Polaromonas naphthalenivorans Species 0.000 description 1
- 108091036407 Polyadenylation Proteins 0.000 description 1
- 239000002202 Polyethylene glycol Substances 0.000 description 1
- 229920002873 Polyethylenimine Polymers 0.000 description 1
- 101710149951 Protein Tat Proteins 0.000 description 1
- 241000590028 Pseudoalteromonas haloplanktis Species 0.000 description 1
- CZPWVGJYEJSRLH-UHFFFAOYSA-N Pyrimidine Chemical compound C1=CN=CN=C1 CZPWVGJYEJSRLH-UHFFFAOYSA-N 0.000 description 1
- 102000014450 RNA Polymerase III Human genes 0.000 description 1
- 108010078067 RNA Polymerase III Proteins 0.000 description 1
- 241000700157 Rattus norvegicus Species 0.000 description 1
- 101001023863 Rattus norvegicus Glucocorticoid receptor Proteins 0.000 description 1
- 101100247004 Rattus norvegicus Qsox1 gene Proteins 0.000 description 1
- 108020004511 Recombinant DNA Proteins 0.000 description 1
- 102000018120 Recombinases Human genes 0.000 description 1
- 108010091086 Recombinases Proteins 0.000 description 1
- 102000006382 Ribonucleases Human genes 0.000 description 1
- 108010083644 Ribonucleases Proteins 0.000 description 1
- 108091028664 Ribonucleotide Proteins 0.000 description 1
- 108020004422 Riboswitch Proteins 0.000 description 1
- 108091006629 SLC13A2 Proteins 0.000 description 1
- 238000012300 Sequence Analysis Methods 0.000 description 1
- 241000191967 Staphylococcus aureus Species 0.000 description 1
- 241000194022 Streptococcus sp. Species 0.000 description 1
- 241001518258 Streptomyces pristinaespiralis Species 0.000 description 1
- 241000272534 Struthio camelus Species 0.000 description 1
- 241000192560 Synechococcus sp. Species 0.000 description 1
- 210000001744 T-lymphocyte Anatomy 0.000 description 1
- 101710137500 T7 RNA polymerase Proteins 0.000 description 1
- 101710192266 Tegument protein VP22 Proteins 0.000 description 1
- 108091046869 Telomeric non-coding RNA Proteins 0.000 description 1
- 241000206213 Thermosipho africanus Species 0.000 description 1
- 241000078013 Trichormus variabilis Species 0.000 description 1
- 108010064978 Type II Site-Specific Deoxyribonucleases Proteins 0.000 description 1
- 108010067022 Type III Site-Specific Deoxyribonucleases Proteins 0.000 description 1
- 108091023045 Untranslated Region Proteins 0.000 description 1
- 208000036142 Viral infection Diseases 0.000 description 1
- 101150095029 W gene Proteins 0.000 description 1
- 210000001766 X chromosome Anatomy 0.000 description 1
- 210000002593 Y chromosome Anatomy 0.000 description 1
- 241001673106 [Bacillus] selenitireducens Species 0.000 description 1
- 210000001015 abdomen Anatomy 0.000 description 1
- 230000005856 abnormality Effects 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 150000007513 acids Chemical class 0.000 description 1
- 230000004913 activation Effects 0.000 description 1
- 210000001789 adipocyte Anatomy 0.000 description 1
- 210000004504 adult stem cell Anatomy 0.000 description 1
- 125000003295 alanine group Chemical group N[C@@H](C)C(=O)* 0.000 description 1
- KOSRFJWDECSPRO-UHFFFAOYSA-N alpha-L-glutamyl-L-glutamic acid Natural products OC(=O)CCC(N)C(=O)NC(CCC(O)=O)C(O)=O KOSRFJWDECSPRO-UHFFFAOYSA-N 0.000 description 1
- 230000003321 amplification Effects 0.000 description 1
- 238000000137 annealing Methods 0.000 description 1
- 239000003242 anti bacterial agent Substances 0.000 description 1
- 229940011019 arthrospira platensis Drugs 0.000 description 1
- 229940009098 aspartate Drugs 0.000 description 1
- 210000003719 b-lymphocyte Anatomy 0.000 description 1
- 238000002869 basic local alignment search tool Methods 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 230000003115 biocidal effect Effects 0.000 description 1
- 230000005540 biological transmission Effects 0.000 description 1
- 229930189065 blasticidin Natural products 0.000 description 1
- 210000002449 bone cell Anatomy 0.000 description 1
- 244000309464 bull Species 0.000 description 1
- 229910000389 calcium phosphate Inorganic materials 0.000 description 1
- 239000001506 calcium phosphate Substances 0.000 description 1
- 235000011010 calcium phosphates Nutrition 0.000 description 1
- 238000004364 calculation method Methods 0.000 description 1
- 210000000748 cardiovascular system Anatomy 0.000 description 1
- 229920006317 cationic polymer Polymers 0.000 description 1
- 150000001768 cations Chemical class 0.000 description 1
- 210000004671 cell-free system Anatomy 0.000 description 1
- 230000010094 cellular senescence Effects 0.000 description 1
- 239000002738 chelating agent Substances 0.000 description 1
- 230000002759 chromosomal effect Effects 0.000 description 1
- 239000011035 citrine Substances 0.000 description 1
- 230000000052 comparative effect Effects 0.000 description 1
- 239000002299 complementary DNA Substances 0.000 description 1
- 238000012790 confirmation Methods 0.000 description 1
- 210000002808 connective tissue Anatomy 0.000 description 1
- 108091036078 conserved sequence Proteins 0.000 description 1
- 101150055601 cops2 gene Proteins 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 230000001186 cumulative effect Effects 0.000 description 1
- 238000005520 cutting process Methods 0.000 description 1
- 210000000805 cytoplasm Anatomy 0.000 description 1
- 239000000412 dendrimer Substances 0.000 description 1
- 229920000736 dendritic polymer Polymers 0.000 description 1
- 239000005547 deoxyribonucleotide Substances 0.000 description 1
- 125000002637 deoxyribonucleotide group Chemical group 0.000 description 1
- 230000001419 dependent effect Effects 0.000 description 1
- 230000004069 differentiation Effects 0.000 description 1
- 210000002249 digestive system Anatomy 0.000 description 1
- 238000006471 dimerization reaction Methods 0.000 description 1
- QNDQILQPPKQROV-UHFFFAOYSA-N dizinc Chemical compound [Zn]=[Zn] QNDQILQPPKQROV-UHFFFAOYSA-N 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 108010057988 ecdysone receptor Proteins 0.000 description 1
- 239000012636 effector Substances 0.000 description 1
- 210000000750 endocrine system Anatomy 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000002255 enzymatic effect Effects 0.000 description 1
- 210000002336 epiblast cell Anatomy 0.000 description 1
- 230000004049 epigenetic modification Effects 0.000 description 1
- 210000002919 epithelial cell Anatomy 0.000 description 1
- 210000000981 epithelium Anatomy 0.000 description 1
- 102000015694 estrogen receptors Human genes 0.000 description 1
- 108010038795 estrogen receptors Proteins 0.000 description 1
- 230000002538 fungal effect Effects 0.000 description 1
- 210000000973 gametocyte Anatomy 0.000 description 1
- 230000002068 genetic effect Effects 0.000 description 1
- 238000010353 genetic engineering Methods 0.000 description 1
- BRZYSWJRSDMWLG-CAXSIQPQSA-N geneticin Natural products O1C[C@@](O)(C)[C@H](NC)[C@@H](O)[C@H]1O[C@@H]1[C@@H](O)[C@H](O[C@@H]2[C@@H]([C@@H](O)[C@H](O)[C@@H](C(C)O)O2)N)[C@@H](N)C[C@H]1N BRZYSWJRSDMWLG-CAXSIQPQSA-N 0.000 description 1
- 108010055341 glutamyl-glutamic acid Proteins 0.000 description 1
- 210000002064 heart cell Anatomy 0.000 description 1
- 210000003494 hepatocyte Anatomy 0.000 description 1
- 230000007062 hydrolysis Effects 0.000 description 1
- 238000006460 hydrolysis reaction Methods 0.000 description 1
- 210000002865 immune cell Anatomy 0.000 description 1
- 238000003119 immunoblot Methods 0.000 description 1
- 238000002513 implantation Methods 0.000 description 1
- 238000007901 in situ hybridization Methods 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 230000000977 initiatory effect Effects 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 238000005304 joining Methods 0.000 description 1
- 210000002510 keratinocyte Anatomy 0.000 description 1
- 229940043355 kinase inhibitor Drugs 0.000 description 1
- 150000002632 lipids Chemical class 0.000 description 1
- 239000002502 liposome Substances 0.000 description 1
- 244000144972 livestock Species 0.000 description 1
- 230000004807 localization Effects 0.000 description 1
- 210000004324 lymphatic system Anatomy 0.000 description 1
- 230000014759 maintenance of location Effects 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 210000002752 melanocyte Anatomy 0.000 description 1
- 229910052751 metal Inorganic materials 0.000 description 1
- 239000002184 metal Substances 0.000 description 1
- 229910021645 metal ion Inorganic materials 0.000 description 1
- 238000000520 microinjection Methods 0.000 description 1
- 230000025608 mitochondrion localization Effects 0.000 description 1
- 108091005601 modified peptides Proteins 0.000 description 1
- 210000001616 monocyte Anatomy 0.000 description 1
- 210000005087 mononuclear cell Anatomy 0.000 description 1
- 210000003205 muscle Anatomy 0.000 description 1
- 210000000663 muscle cell Anatomy 0.000 description 1
- 230000003387 muscular Effects 0.000 description 1
- CJWXCNXHAIFFMH-AVZHFPDBSA-N n-[(2s,3r,4s,5s,6r)-2-[(2r,3r,4s,5r)-2-acetamido-4,5,6-trihydroxy-1-oxohexan-3-yl]oxy-3,5-dihydroxy-6-methyloxan-4-yl]acetamide Chemical compound C[C@H]1O[C@@H](O[C@@H]([C@@H](O)[C@H](O)CO)[C@@H](NC(C)=O)C=O)[C@H](O)[C@@H](NC(C)=O)[C@@H]1O CJWXCNXHAIFFMH-AVZHFPDBSA-N 0.000 description 1
- 239000002105 nanoparticle Substances 0.000 description 1
- 230000006780 non-homologous end joining Effects 0.000 description 1
- 238000003199 nucleic acid amplification method Methods 0.000 description 1
- 125000000371 nucleobase group Chemical group 0.000 description 1
- 230000003287 optical effect Effects 0.000 description 1
- 210000003463 organelle Anatomy 0.000 description 1
- 210000003101 oviduct Anatomy 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 230000037361 pathway Effects 0.000 description 1
- 230000035515 penetration Effects 0.000 description 1
- 230000026731 phosphorylation Effects 0.000 description 1
- 238000006366 phosphorylation reaction Methods 0.000 description 1
- 239000003757 phosphotransferase inhibitor Substances 0.000 description 1
- 230000004962 physiological condition Effects 0.000 description 1
- 239000013600 plasmid vector Substances 0.000 description 1
- 230000008488 polyadenylation Effects 0.000 description 1
- 108010011110 polyarginine Proteins 0.000 description 1
- 229920001223 polyethylene glycol Polymers 0.000 description 1
- 102000054765 polymorphisms of proteins Human genes 0.000 description 1
- 230000029279 positive regulation of transcription, DNA-dependent Effects 0.000 description 1
- 210000000229 preadipocyte Anatomy 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 230000035755 proliferation Effects 0.000 description 1
- 230000007026 protein scission Effects 0.000 description 1
- 230000004850 protein–protein interaction Effects 0.000 description 1
- 229950010131 puromycin Drugs 0.000 description 1
- 238000011002 quantification Methods 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 102000037983 regulatory factors Human genes 0.000 description 1
- 108091008025 regulatory factors Proteins 0.000 description 1
- 230000008439 repair process Effects 0.000 description 1
- 210000004994 reproductive system Anatomy 0.000 description 1
- 210000002345 respiratory system Anatomy 0.000 description 1
- 230000004044 response Effects 0.000 description 1
- 230000000717 retained effect Effects 0.000 description 1
- 239000002336 ribonucleotide Substances 0.000 description 1
- 125000002652 ribonucleotide group Chemical group 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 230000003584 silencer Effects 0.000 description 1
- 238000002741 site-directed mutagenesis Methods 0.000 description 1
- 210000004683 skeletal myoblast Anatomy 0.000 description 1
- 210000000329 smooth muscle myocyte Anatomy 0.000 description 1
- 238000003153 stable transfection Methods 0.000 description 1
- 150000003431 steroids Chemical class 0.000 description 1
- 238000010809 targeting technique Methods 0.000 description 1
- 101150075675 tatC gene Proteins 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 230000005026 transcription initiation Effects 0.000 description 1
- 230000005030 transcription termination Effects 0.000 description 1
- 108091006106 transcriptional activators Proteins 0.000 description 1
- 238000003146 transient transfection Methods 0.000 description 1
- 230000007704 transition Effects 0.000 description 1
- 238000013519 translation Methods 0.000 description 1
- 230000014621 translational initiation Effects 0.000 description 1
- QORWJWZARLRLPR-UHFFFAOYSA-H tricalcium bis(phosphate) Chemical compound [Ca+2].[Ca+2].[Ca+2].[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O QORWJWZARLRLPR-UHFFFAOYSA-H 0.000 description 1
- GWBUNZLLLLDXMD-UHFFFAOYSA-H tricopper;dicarbonate;dihydroxide Chemical compound [OH-].[OH-].[Cu+2].[Cu+2].[Cu+2].[O-]C([O-])=O.[O-]C([O-])=O GWBUNZLLLLDXMD-UHFFFAOYSA-H 0.000 description 1
- 241000701161 unidentified adenovirus Species 0.000 description 1
- 241001430294 unidentified retrovirus Species 0.000 description 1
- 230000002485 urinary effect Effects 0.000 description 1
- 230000009385 viral infection Effects 0.000 description 1
- 230000003612 virological effect Effects 0.000 description 1
- 230000001018 virulence Effects 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/87—Introduction of foreign genetic material using processes not otherwise provided for, e.g. co-transformation
- C12N15/90—Stable introduction of foreign DNA into chromosome
- C12N15/902—Stable introduction of foreign DNA into chromosome using homologous recombination
- C12N15/907—Stable introduction of foreign DNA into chromosome using homologous recombination in mammalian cells
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K67/00—Rearing or breeding animals, not otherwise provided for; New or modified breeds of animals
- A01K67/027—New or modified breeds of vertebrates
- A01K67/0275—Genetically modified vertebrates, e.g. transgenic
- A01K67/0276—Knock-out vertebrates
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K67/00—Rearing or breeding animals, not otherwise provided for; New or modified breeds of animals
- A01K67/027—New or modified breeds of vertebrates
- A01K67/0275—Genetically modified vertebrates, e.g. transgenic
- A01K67/0278—Knock-in vertebrates, e.g. humanised vertebrates
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/14—Hydrolases (3)
- C12N9/16—Hydrolases (3) acting on ester bonds (3.1)
- C12N9/22—Ribonucleases RNAses, DNAses
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K2217/00—Genetically modified animals
- A01K2217/07—Animals genetically altered by homologous recombination
- A01K2217/072—Animals genetically altered by homologous recombination maintaining or altering function, i.e. knock in
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K2217/00—Genetically modified animals
- A01K2217/07—Animals genetically altered by homologous recombination
- A01K2217/075—Animals genetically altered by homologous recombination inducing loss of function, i.e. knock out
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K2227/00—Animals characterised by species
- A01K2227/10—Mammal
- A01K2227/105—Murine
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
- C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
- C12N15/85—Vectors or expression systems specially adapted for eukaryotic hosts for animal cells
- C12N15/8509—Vectors or expression systems specially adapted for eukaryotic hosts for animal cells for producing genetically modified animals, e.g. transgenic
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Engineering & Computer Science (AREA)
- Zoology (AREA)
- Biotechnology (AREA)
- Chemical & Material Sciences (AREA)
- Wood Science & Technology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Organic Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Molecular Biology (AREA)
- Biomedical Technology (AREA)
- General Engineering & Computer Science (AREA)
- Environmental Sciences (AREA)
- Veterinary Medicine (AREA)
- Biochemistry (AREA)
- Microbiology (AREA)
- Animal Behavior & Ethology (AREA)
- Biodiversity & Conservation Biology (AREA)
- Animal Husbandry (AREA)
- Physics & Mathematics (AREA)
- Plant Pathology (AREA)
- Biophysics (AREA)
- Cell Biology (AREA)
- Mycology (AREA)
- Medicinal Chemistry (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Enzymes And Modification Thereof (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
Abstract
Cette invention concerne des procédés et des compositions permettant de réaliser une ou plusieurs modifications génétiques ciblées au niveau d'un locus génomique cible à l'intérieur d'une cellule et de produire des animaux non humains comprenant le locus génomique modifié. Ces procédés font appel à deux ou plusieurs grands vecteurs de ciblage aptes à se recombiner les uns avec les autres et avec le locus génomique cible dans une seule étape de ciblage génomique. Ces procédés peuvent également être utilisés en combinaison avec un agent de nucléase. L'invention concerne également des procédés et des compositions fournis pour améliorer la recombinaison homologue au niveau d'un locus génomique cible dans une cellule. Ces procédés consistent à utiliser au moins deux acides nucléiques comprenant une ou plusieurs séquences de chevauchement. Ces procédés peuvent être utilisés en combinaison avec un agent de nucléase ou sans un agent de nucléase.
Applications Claiming Priority (7)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US201462094104P | 2014-12-19 | 2014-12-19 | |
| US62/094,104 | 2014-12-19 | ||
| US201562167408P | 2015-05-28 | 2015-05-28 | |
| US62/167,408 | 2015-05-28 | ||
| US201562205524P | 2015-08-14 | 2015-08-14 | |
| US62/205,524 | 2015-08-14 | ||
| PCT/US2015/066681 WO2016100819A1 (fr) | 2014-12-19 | 2015-12-18 | Procédés et compositions pour modification génétique ciblée par ciblage multiple en une seule étape |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CA2971213A1 CA2971213A1 (fr) | 2016-06-23 |
| CA2971213C true CA2971213C (fr) | 2023-09-26 |
Family
ID=55229831
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CA2971213A Active CA2971213C (fr) | 2014-12-19 | 2015-12-18 | Procedes et compositions pour modification genetique ciblee par ciblage multiple en une seule etape |
Country Status (15)
| Country | Link |
|---|---|
| US (2) | US11326184B2 (fr) |
| EP (2) | EP3653048B9 (fr) |
| JP (2) | JP6840077B2 (fr) |
| KR (1) | KR102530821B1 (fr) |
| CN (1) | CN107208113A (fr) |
| AU (1) | AU2015364427B2 (fr) |
| BR (1) | BR112017013104A2 (fr) |
| CA (1) | CA2971213C (fr) |
| ES (2) | ES2947714T3 (fr) |
| IL (1) | IL252755B (fr) |
| MX (1) | MX2017008190A (fr) |
| NZ (1) | NZ732895A (fr) |
| RU (1) | RU2707137C2 (fr) |
| SG (1) | SG11201704646YA (fr) |
| WO (1) | WO2016100819A1 (fr) |
Families Citing this family (26)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2013163394A1 (fr) | 2012-04-25 | 2013-10-31 | Regeneron Pharmaceuticals, Inc. | Ciblage médié par nucléase avec de grands vecteurs de ciblage |
| JP6411463B2 (ja) | 2013-04-16 | 2018-10-24 | リジェネロン・ファーマシューティカルズ・インコーポレイテッドＲｅｇｅｎｅｒｏｎ Ｐｈａｒｍａｃｅｕｔｉｃａｌｓ， Ｉｎｃ． | ラットゲノムの標的改変 |
| RU2685914C1 (ru) | 2013-12-11 | 2019-04-23 | Регенерон Фармасьютикалс, Инк. | Способы и композиции для направленной модификации генома |
| PL3152312T3 (pl) | 2014-06-06 | 2020-08-10 | Regeneron Pharmaceuticals, Inc. | Sposoby i kompozycje do modyfikowania docelowego locus |
| MX2016017317A (es) | 2014-06-26 | 2017-08-24 | Regeneron Pharma | Metodos y composiciones para modificaciones geneticas dirigidas y metodos de uso. |
| US10428310B2 (en) | 2014-10-15 | 2019-10-01 | Regeneron Pharmaceuticals, Inc. | Methods and compositions for generating or maintaining pluripotent cells |
| US11680268B2 (en) | 2014-11-07 | 2023-06-20 | Editas Medicine, Inc. | Methods for improving CRISPR/Cas-mediated genome-editing |
| KR102531016B1 (ko) | 2014-11-21 | 2023-05-10 | 리제너론 파마슈티칼스 인코포레이티드 | 쌍 형성된 가이드 rna를 사용하는 표적화된 유전자 변형을 위한 방법 및 조성물 |
| RU2707137C2 (ru) * | 2014-12-19 | 2019-11-22 | Регенерон Фармасьютикалз, Инк. | Способы и композиции для нацеленной генетической модификации посредством одноэтапного множественного нацеливания |
| US10285388B2 (en) | 2015-05-29 | 2019-05-14 | Regeneron Pharmaceuticals, Inc. | Non-human animals having a disruption in a C9ORF72 locus |
| AU2016326711B2 (en) | 2015-09-24 | 2022-11-03 | Editas Medicine, Inc. | Use of exonucleases to improve CRISPR/Cas-mediated genome editing |
| MX2018008701A (es) | 2016-01-13 | 2019-06-06 | Regeneron Pharma | Roedores que tienen una region de diversidad de cadena pesada modificada geneticamente. |
| EP3433363A1 (fr) | 2016-03-25 | 2019-01-30 | Editas Medicine, Inc. | Systèmes d'édition de génome comprenant des molécules d'enzyme modulant la réparation et leurs procédés d'utilisation |
| US11236313B2 (en) | 2016-04-13 | 2022-02-01 | Editas Medicine, Inc. | Cas9 fusion molecules, gene editing systems, and methods of use thereof |
| AU2017336100B2 (en) | 2016-09-30 | 2023-11-30 | Regeneron Pharmaceuticals, Inc. | Non-human animals having a hexanucleotide repeat expansion in a C9ORF72 locus |
| DK3766343T3 (da) | 2016-11-04 | 2022-06-20 | Regeneron Pharma | Ikke-humane dyr med et manipuleret immunoglobulin-lambda-letkædelocus |
| IL267470B2 (en) * | 2017-01-10 | 2023-10-01 | F O R E Biotherapeutics Ltd | Methods for in vitro site-directed mutagenesis using gene editing technologies |
| WO2019014564A1 (fr) | 2017-07-14 | 2019-01-17 | Editas Medicine, Inc. | Systèmes et procédés d'intégration ciblée et d'édition du génome et détection de celle-ci à l'aide de sites d'amorçage intégrés |
| JP2020533957A (ja) * | 2017-07-31 | 2020-11-26 | リジェネロン・ファーマシューティカルズ・インコーポレイテッドＲｅｇｅｎｅｒｏｎ Ｐｈａｒｍａｃｅｕｔｉｃａｌｓ， Ｉｎｃ． | Ｃｒｉｓｐｒリポーター非ヒト動物およびその使用 |
| CA3084049A1 (fr) | 2017-12-05 | 2019-06-13 | Regeneron Pharmaceuticals, Inc. | Animaux non humains presentant une chaine legere d'immunoglobuline lambda modifiee et leurs utilisations |
| SI3629720T1 (sl) | 2018-06-14 | 2022-04-29 | Regeneron Pharmaceuticals, Inc. | Nehumane živali zmožne prerazporeditve DH-DH v kodirnih zaporedjih težke verige imunoglobulina |
| CN111321171A (zh) * | 2018-12-14 | 2020-06-23 | 江苏集萃药康生物科技有限公司 | 一种应用CRISPR/Cas9介导ES打靶技术制备基因打靶动物模型的方法 |
| JP7449291B2 (ja) | 2018-12-20 | 2024-03-13 | リジェネロン・ファーマシューティカルズ・インコーポレイテッド | ヌクレアーゼ媒介リピート伸長 |
| CN113795588A (zh) * | 2019-04-04 | 2021-12-14 | 瑞泽恩制药公司 | 用于在靶向性载体中无瘢痕引入靶向修饰的方法 |
| TW202110323A (zh) | 2019-06-05 | 2021-03-16 | 美商再生元醫藥公司 | 具有表現自κ 基因座的有限λ 輕鏈組庫的非人類動物及其用途 |
| US20220195014A1 (en) | 2020-12-23 | 2022-06-23 | Regeneron Pharmaceuticals, Inc. | Nucleic acids encoding anchor modified antibodies and uses thereof |
Family Cites Families (94)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0546091B1 (fr) * | 1990-08-29 | 2007-01-24 | Pharming Intellectual Property BV | Recombinaison homologue dans des cellules de mammiferes |
| US5830729A (en) | 1996-04-18 | 1998-11-03 | Institut Pasteur | I Sce I-induced gene replacement and gene conversion in embryonic stem cells |
| AU8587598A (en) | 1997-07-26 | 1999-02-16 | Wisconsin Alumni Research Foundation | Trans-species nuclear transfer |
| WO2000039316A1 (fr) | 1998-12-31 | 2000-07-06 | The J. David Gladstone Institutes | Rongeurs transgeniques et lignees cellulaires de rongeur exprimant des co-recepteurs du vih |
| US6599692B1 (en) | 1999-09-14 | 2003-07-29 | Sangamo Bioscience, Inc. | Functional genomics using zinc finger proteins |
| US20030104526A1 (en) | 1999-03-24 | 2003-06-05 | Qiang Liu | Position dependent recognition of GNN nucleotide triplets by zinc fingers |
| ATE309536T1 (de) | 1999-12-06 | 2005-11-15 | Sangamo Biosciences Inc | Methoden zur verwendung von randomisierten zinkfingerprotein-bibliotheken zur identifizierung von genfunktionen |
| US20050144655A1 (en) | 2000-10-31 | 2005-06-30 | Economides Aris N. | Methods of modifying eukaryotic cells |
| US7105348B2 (en) | 2000-10-31 | 2006-09-12 | Regeneron Pharmaceuticals, Inc. | Methods of modifying eukaryotic cells |
| US6596541B2 (en) | 2000-10-31 | 2003-07-22 | Regeneron Pharmaceuticals, Inc. | Methods of modifying eukaryotic cells |
| US6586251B2 (en) * | 2000-10-31 | 2003-07-01 | Regeneron Pharmaceuticals, Inc. | Methods of modifying eukaryotic cells |
| AU2884102A (en) | 2000-12-07 | 2002-06-18 | Sangamo Biosciences Inc | Regulation of angiogenesis with zinc finger proteins |
| AU2002225187A1 (en) | 2001-01-22 | 2002-07-30 | Sangamo Biosciences, Inc. | Zinc finger polypeptides and their use |
| CA2435394C (fr) | 2001-01-22 | 2018-01-09 | Sangamo Biosciences, Inc. | Proteines de liaison en doigt de gant modifiees |
| AUPR451401A0 (en) | 2001-04-20 | 2001-05-24 | Monash University | A method of nuclear transfer |
| AU2003251286B2 (en) | 2002-01-23 | 2007-08-16 | The University Of Utah Research Foundation | Targeted chromosomal mutagenesis using zinc finger nucleases |
| US20060078552A1 (en) | 2002-03-15 | 2006-04-13 | Sylvain Arnould | Hybrid and single chain meganucleases and use thereof |
| US20030232410A1 (en) | 2002-03-21 | 2003-12-18 | Monika Liljedahl | Methods and compositions for using zinc finger endonucleases to enhance homologous recombination |
| US7612250B2 (en) | 2002-07-29 | 2009-11-03 | Trustees Of Tufts College | Nuclear transfer embryo formation method |
| CA2497913C (fr) | 2002-09-05 | 2014-06-03 | California Institute Of Technology | Utilisation de nucleases chimeres pour stimuler le ciblage de genes |
| AU2003290518A1 (en) | 2002-09-06 | 2004-04-23 | Fred Hutchinson Cancer Research Center | Methods and compositions concerning designed highly-specific nucleic acid binding proteins |
| US20030175968A1 (en) | 2002-10-30 | 2003-09-18 | Golic Kent G. | Gene targeting method |
| AU2004204509A1 (en) | 2003-01-13 | 2004-07-29 | Mario R. Capecchi | Persistent expression of candidate molecule in proliferating stem and progenitor cells for delivery of therapeutic products |
| US7888121B2 (en) | 2003-08-08 | 2011-02-15 | Sangamo Biosciences, Inc. | Methods and compositions for targeted cleavage and recombination |
| US8409861B2 (en) | 2003-08-08 | 2013-04-02 | Sangamo Biosciences, Inc. | Targeted deletion of cellular DNA sequences |
| EP1591521A1 (fr) | 2004-04-30 | 2005-11-02 | Cellectis | Dérivés de I-Dmo I ayant une activité améliorée à 37degrés C et leur utilisation |
| CA2579677A1 (fr) | 2004-09-16 | 2006-03-30 | Sangamo Biosciences, Inc. | Production de proteines: compositions et methodes |
| DK2767161T3 (en) | 2004-10-19 | 2018-05-07 | Regeneron Pharma | Method of generating an animal homozygous for genetic modification |
| FR2879622B1 (fr) | 2004-12-17 | 2008-02-01 | Agronomique Inst Nat Rech | Procede in vitro de production d'ovocytes ou d'oeufs presentant une modification genomique ciblee |
| WO2006097784A1 (fr) | 2005-03-15 | 2006-09-21 | Cellectis | Variants de meganuclease i-crei presentant une specificite modifiee, leur procede de preparation, et leurs utilisations |
| AU2006224248B2 (en) | 2005-03-15 | 2011-01-06 | Cellectis | I-Crei meganuclease variants with modified specificity, method of preparation and uses thereof |
| GB0615327D0 (en) | 2006-03-30 | 2006-09-13 | Univ Edinburgh | Culture medium containing kinase inhibitors and uses thereof |
| WO2007117410A2 (fr) | 2006-03-31 | 2007-10-18 | Medarex, Inc. | Animaux transgéniques exprimant des anticorps chimériques destinés à être utilisés pour la préparation d'anticorps humains |
| CN101117633B (zh) | 2006-08-03 | 2011-07-20 | 上海交通大学附属儿童医院 | 一种细胞核移植方法 |
| JP5632610B2 (ja) | 2006-12-14 | 2014-11-26 | ダウ アグロサイエンシィズ エルエルシー | 最適化された非正準ジンクフィンガータンパク質 |
| US7771967B2 (en) | 2006-12-22 | 2010-08-10 | The J. David Gladstone Institutes | Nucleic acid encoding apolipoprotein E-I3 |
| WO2008133938A2 (fr) | 2007-04-26 | 2008-11-06 | Sangamo Biosciences, Inc. | Intégration ciblée dans le locus ppp1r12c |
| CN101784664B (zh) | 2007-06-01 | 2013-04-10 | Omt公司 | 用于抑制内源性免疫球蛋白基因和生产转基因人独特型抗体的组合物和方法 |
| GB0803109D0 (en) * | 2008-02-20 | 2008-03-26 | Gene Bridges Gmbh | Method of nucleic acid recombination |
| KR101803737B1 (ko) | 2008-12-04 | 2017-12-01 | 상가모 테라퓨틱스, 인코포레이티드 | 징크 핑거 뉴클레아제를 이용한 랫트의 유전체 편집 |
| US20110239315A1 (en) | 2009-01-12 | 2011-09-29 | Ulla Bonas | Modular dna-binding domains and methods of use |
| EP2206723A1 (fr) | 2009-01-12 | 2010-07-14 | Bonas, Ulla | Domaines modulaires de liaison à l'ADN |
| JP5932632B2 (ja) | 2009-03-20 | 2016-06-15 | サンガモ バイオサイエンシーズ， インコーポレイテッド | 改変された亜鉛フィンガータンパク質を使用したｃｘｃｒ４の修飾 |
| US8772008B2 (en) | 2009-05-18 | 2014-07-08 | Sangamo Biosciences, Inc. | Methods and compositions for increasing nuclease activity |
| US20120178647A1 (en) | 2009-08-03 | 2012-07-12 | The General Hospital Corporation | Engineering of zinc finger arrays by context-dependent assembly |
| WO2011051390A1 (fr) | 2009-10-28 | 2011-05-05 | Helmholtz Zentrum München - Deutsches Forschungszentrum für Gesundheit und Umwelt (GmbH) | Recombinaison homologue dans l'ovocyte |
| CA2779858C (fr) | 2009-10-29 | 2019-10-29 | Aris N. Economides | Alleles multifonctionnels |
| US20120315670A1 (en) | 2009-11-02 | 2012-12-13 | Gen9, Inc. | Compositions and Methods for the Regulation of Multiple Genes of Interest in a Cell |
| JP5807862B2 (ja) | 2009-12-01 | 2015-11-10 | 国立研究開発法人国立がん研究センター | ラット胚性幹細胞を用いたキメララットの作製法 |
| WO2011072246A2 (fr) | 2009-12-10 | 2011-06-16 | Regents Of The University Of Minnesota | Modification de l'adn induite par l'effecteur tal |
| JP5924773B2 (ja) | 2009-12-21 | 2016-05-25 | キージーン・エン・フェー | プロトプラストを形質移入するための改善された技術 |
| EP2660318A1 (fr) | 2010-02-09 | 2013-11-06 | Sangamo BioSciences, Inc. | Modification génomique ciblée de molécules partiellement donneuses à simple brin |
| GB201009732D0 (en) * | 2010-06-10 | 2010-07-21 | Gene Bridges Gmbh | Direct cloning |
| CA2800534C (fr) | 2010-06-11 | 2019-01-22 | Regeneron Pharmaceuticals, Inc. | Production d'animaux femelles xy fertiles a partir de cellules es xy |
| CA2993567C (fr) | 2010-07-21 | 2022-06-28 | Sangamo Biosciences, Inc. | Methodes et compositions destinees a la modification d'un gene recepteurde lymphocyte t |
| BR112013001685B1 (pt) | 2010-07-23 | 2021-10-13 | Sigma-Aldrich Co. Llc | Método para editar pelo menos uma sequência cromossômica endógena em uma célula pela inserção de uma sequência na sequência cromossômica |
| WO2012018726A1 (fr) | 2010-08-02 | 2012-02-09 | Cellectis Sa | Procédé d'augmentation du ciblage de gènes à cassure double-brin induite |
| WO2012129198A1 (fr) | 2011-03-23 | 2012-09-27 | Transposagen Biopharmaceuticals, Inc. | Modèles de rat génétiquement modifiés pour l'étude de l'obésité et du diabète |
| AU2012327204B2 (en) | 2011-10-28 | 2016-01-21 | Regeneron Pharmaceuticals, Inc. | Genetically modified T cell receptor mice |
| US9637739B2 (en) | 2012-03-20 | 2017-05-02 | Vilnius University | RNA-directed DNA cleavage by the Cas9-crRNA complex |
| WO2013141680A1 (fr) | 2012-03-20 | 2013-09-26 | Vilnius University | Clivage d'adn dirigé par arn par le complexe cas9-arncr |
| WO2013163394A1 (fr) * | 2012-04-25 | 2013-10-31 | Regeneron Pharmaceuticals, Inc. | Ciblage médié par nucléase avec de grands vecteurs de ciblage |
| CN104471067B (zh) | 2012-05-07 | 2020-08-14 | 桑格摩生物治疗股份有限公司 | 用于核酸酶介导的转基因靶向整合的方法和组合物 |
| PT2800811T (pt) | 2012-05-25 | 2017-08-17 | Univ California | Métodos e composições para modificação de adn alvo dirigida por arn e para modulação dirigida por arn de transcrição |
| CN110643600A (zh) | 2012-10-23 | 2020-01-03 | 基因工具股份有限公司 | 用于切割靶dna的系统及其用途 |
| ES2769310T3 (es) | 2012-12-06 | 2020-06-25 | Sigma Aldrich Co Llc | Modificación y regulación del genoma basada en CRISPR |
| MX2015007550A (es) | 2012-12-12 | 2017-02-02 | Broad Inst Inc | Suministro, modificación y optimización de sistemas, métodos y composiciones para la manipulación de secuencias y aplicaciones terapéuticas. |
| US8697359B1 (en) | 2012-12-12 | 2014-04-15 | The Broad Institute, Inc. | CRISPR-Cas systems and methods for altering expression of gene products |
| SG10201912327SA (en) | 2012-12-12 | 2020-02-27 | Broad Inst Inc | Engineering and Optimization of Improved Systems, Methods and Enzyme Compositions for Sequence Manipulation |
| PT3653049T (pt) * | 2012-12-14 | 2023-11-14 | Omniab Inc | Polinucleótidos que codificam anticorpos de roedores com idiótipos humanos e animais que os compreendem |
| EP3553174A1 (fr) | 2012-12-17 | 2019-10-16 | President and Fellows of Harvard College | Ingénierie de génome humain guidée par arn |
| JP6583918B2 (ja) | 2012-12-27 | 2019-10-02 | キージーン ナムローゼ フェンノートシャップ | 植物における遺伝連鎖を解消するための方法 |
| RU2690352C2 (ru) | 2013-02-20 | 2019-05-31 | Регенерон Фармасютикалс, Инк. | Генетическая модификация крыс |
| JP2016507244A (ja) | 2013-02-27 | 2016-03-10 | ヘルムホルツ・ツェントルム・ミュンヒェン・ドイチェス・フォルシュンクスツェントルム・フューア・ゲズントハイト・ウント・ウムベルト（ゲーエムベーハー）Ｈｅｌｍｈｏｌｔｚ Ｚｅｎｔｒｕｍ ＭｕｅｎｃｈｅｎＤｅｕｔｓｃｈｅｓ Ｆｏｒｓｃｈｕｎｇｓｚｅｎｔｒｕｍ ｆｕｅｒ Ｇｅｓｕｎｄｈｅｉｔ ｕｎｄ Ｕｍｗｅｌｔ （ＧｍｂＨ） | Ｃａｓ９ヌクレアーゼによる卵母細胞における遺伝子編集 |
| US10612043B2 (en) | 2013-03-09 | 2020-04-07 | Agilent Technologies, Inc. | Methods of in vivo engineering of large sequences using multiple CRISPR/cas selections of recombineering events |
| US9234213B2 (en) | 2013-03-15 | 2016-01-12 | System Biosciences, Llc | Compositions and methods directed to CRISPR/Cas genomic engineering systems |
| EP2975942B1 (fr) | 2013-03-21 | 2018-08-08 | Sangamo Therapeutics, Inc. | Interruption ciblée de gènes de récepteur des lymphocytes t à l'aide de nucléases protéiques à doigt de zinc génétiquement modifiées |
| JP6411463B2 (ja) * | 2013-04-16 | 2018-10-24 | リジェネロン・ファーマシューティカルズ・インコーポレイテッドＲｅｇｅｎｅｒｏｎ Ｐｈａｒｍａｃｅｕｔｉｃａｌｓ， Ｉｎｃ． | ラットゲノムの標的改変 |
| WO2015040402A1 (fr) | 2013-09-18 | 2015-03-26 | Kymab Limited | Procédés, cellules et organismes |
| US10787684B2 (en) | 2013-11-19 | 2020-09-29 | President And Fellows Of Harvard College | Large gene excision and insertion |
| RU2685914C1 (ru) | 2013-12-11 | 2019-04-23 | Регенерон Фармасьютикалс, Инк. | Способы и композиции для направленной модификации генома |
| US9476065B2 (en) | 2013-12-19 | 2016-10-25 | Amyris, Inc. | Methods for genomic integration |
| KR101823661B1 (ko) | 2014-04-24 | 2018-01-30 | 기초과학연구원 | 마이크로 상동 기반의 유전자 녹아웃을 위한 뉴클레아제 표적 서열을 확인하는 방법 |
| GB201407852D0 (en) | 2014-05-02 | 2014-06-18 | Iontas Ltd | Preparation of libraries od protein variants expressed in eukaryotic cells and use for selecting binding molecules |
| PL3152312T3 (pl) | 2014-06-06 | 2020-08-10 | Regeneron Pharmaceuticals, Inc. | Sposoby i kompozycje do modyfikowania docelowego locus |
| MX2016017317A (es) | 2014-06-26 | 2017-08-24 | Regeneron Pharma | Metodos y composiciones para modificaciones geneticas dirigidas y metodos de uso. |
| CA2959428A1 (fr) * | 2014-09-19 | 2016-03-24 | Regeneron Pharmaceuticals, Inc. | Recepteurs d'antigenes chimeriques |
| WO2016061073A1 (fr) | 2014-10-14 | 2016-04-21 | Memorial Sloan-Kettering Cancer Center | Composition et technique d'ingénierie in vivo pour réarrangements chromosomiques |
| US10428310B2 (en) | 2014-10-15 | 2019-10-01 | Regeneron Pharmaceuticals, Inc. | Methods and compositions for generating or maintaining pluripotent cells |
| JP6772067B2 (ja) | 2014-11-20 | 2020-10-21 | 国立大学法人京都大学 | 哺乳動物の標的ゲノム領域にｄｎａをノックインする方法及び細胞 |
| KR102531016B1 (ko) | 2014-11-21 | 2023-05-10 | 리제너론 파마슈티칼스 인코포레이티드 | 쌍 형성된 가이드 rna를 사용하는 표적화된 유전자 변형을 위한 방법 및 조성물 |
| US20170266320A1 (en) | 2014-12-01 | 2017-09-21 | President And Fellows Of Harvard College | RNA-Guided Systems for In Vivo Gene Editing |
| RU2707137C2 (ru) | 2014-12-19 | 2019-11-22 | Регенерон Фармасьютикалз, Инк. | Способы и композиции для нацеленной генетической модификации посредством одноэтапного множественного нацеливания |
| US9790490B2 (en) | 2015-06-18 | 2017-10-17 | The Broad Institute Inc. | CRISPR enzymes and systems |
-
2015
- 2015-12-18 RU RU2017124909A patent/RU2707137C2/ru active
- 2015-12-18 NZ NZ732895A patent/NZ732895A/en unknown
- 2015-12-18 ES ES19201384T patent/ES2947714T3/es active Active
- 2015-12-18 WO PCT/US2015/066681 patent/WO2016100819A1/fr active Application Filing
- 2015-12-18 SG SG11201704646YA patent/SG11201704646YA/en unknown
- 2015-12-18 US US14/974,623 patent/US11326184B2/en active Active
- 2015-12-18 AU AU2015364427A patent/AU2015364427B2/en active Active
- 2015-12-18 EP EP19201384.5A patent/EP3653048B9/fr active Active
- 2015-12-18 MX MX2017008190A patent/MX2017008190A/es unknown
- 2015-12-18 CN CN201580069507.5A patent/CN107208113A/zh active Pending
- 2015-12-18 BR BR112017013104A patent/BR112017013104A2/pt not_active Application Discontinuation
- 2015-12-18 JP JP2017532808A patent/JP6840077B2/ja active Active
- 2015-12-18 EP EP15828604.7A patent/EP3232774B1/fr active Active
- 2015-12-18 ES ES15828604T patent/ES2760508T3/es active Active
- 2015-12-18 KR KR1020177019826A patent/KR102530821B1/ko active IP Right Grant
- 2015-12-18 CA CA2971213A patent/CA2971213C/fr active Active
-
2017
- 2017-06-07 IL IL252755A patent/IL252755B/en unknown
-
2020
- 2020-12-28 JP JP2020219029A patent/JP7095066B2/ja active Active
-
2022
- 2022-04-05 US US17/713,696 patent/US20220235381A1/en active Pending
Also Published As
| Publication number | Publication date |
|---|---|
| CA2971213A1 (fr) | 2016-06-23 |
| ES2760508T3 (es) | 2020-05-14 |
| US20220235381A1 (en) | 2022-07-28 |
| IL252755A0 (en) | 2017-08-31 |
| EP3653048B9 (fr) | 2023-10-04 |
| AU2015364427A1 (en) | 2017-07-06 |
| KR102530821B1 (ko) | 2023-05-10 |
| US11326184B2 (en) | 2022-05-10 |
| EP3653048A1 (fr) | 2020-05-20 |
| RU2707137C2 (ru) | 2019-11-22 |
| WO2016100819A1 (fr) | 2016-06-23 |
| BR112017013104A2 (pt) | 2018-05-15 |
| EP3232774B1 (fr) | 2019-10-09 |
| CN107208113A (zh) | 2017-09-26 |
| JP2021045174A (ja) | 2021-03-25 |
| NZ732895A (en) | 2022-05-27 |
| EP3232774A1 (fr) | 2017-10-25 |
| JP7095066B2 (ja) | 2022-07-04 |
| IL252755B (en) | 2021-10-31 |
| ES2947714T3 (es) | 2023-08-17 |
| MX2017008190A (es) | 2018-03-23 |
| SG11201704646YA (en) | 2017-07-28 |
| RU2017124909A (ru) | 2019-01-21 |
| KR20170093246A (ko) | 2017-08-14 |
| AU2015364427B2 (en) | 2021-05-27 |
| JP2017538428A (ja) | 2017-12-28 |
| ES2947714T9 (es) | 2024-03-14 |
| RU2017124909A3 (fr) | 2019-06-05 |
| US20160177339A1 (en) | 2016-06-23 |
| EP3653048C0 (fr) | 2023-06-07 |
| JP6840077B2 (ja) | 2021-03-10 |
| EP3653048B1 (fr) | 2023-06-07 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| US20220235381A1 (en) | Methods and compositions for targeted genetic modification through single-step multiple targeting | |
| AU2021201239B2 (en) | Methods and compositions for targeted genetic modifications and methods of use | |
| US10294494B2 (en) | Methods and compositions for modifying a targeted locus |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| EEER | Examination request |
Effective date: 20201211 |
|
| EEER | Examination request |
Effective date: 20201211 |
|
| EEER | Examination request |
Effective date: 20201211 |
|
| EEER | Examination request |
Effective date: 20201211 |
|
| EEER | Examination request |
Effective date: 20201211 |
|
| EEER | Examination request |
Effective date: 20201211 |
|
| EEER | Examination request |
Effective date: 20201211 |
|
| EEER | Examination request |
Effective date: 20201211 |